Difference between revisions of "Part:BBa K4047024"
 
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<partinfo>BBa_K4047024 short</partinfo>
 
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The backbone sequence used for restriction digest assembly of our pGEM-FBPase, pGEM-tal, and pGEM-Tal/FBPase plasmids (<partinfo>BBa_K4047036</partinfo>, <partinfo>BBa_K4047037</partinfo>, <partinfo>BBa_K4047038</partinfo>). Its multiple components include an origin of replication (<partinfo>BBa_K4047029</partinfo>), a trc promoter (<partinfo>BBa_K4047032</partinfo>), an rrnB T1 terminator (<partinfo>BBa_K4047025</partinfo>), an rrnB T2 terminator (<partinfo>BBa_K4047026</partinfo>), a base of mobility region (<partinfo>BBa_K4047028</partinfo>), a lac repressor (<partinfo>BBa_K4047031</partinfo>), a lacIq promoter (<partinfo>BBa_K4047030</partinfo>), a lac operator (<partinfo>BBa_K4047033</partinfo>), and spectinomycin/streptomycin resistance (<partinfo>BBa_K4047027</partinfo>).  
 
The backbone sequence used for restriction digest assembly of our pGEM-FBPase, pGEM-tal, and pGEM-Tal/FBPase plasmids (<partinfo>BBa_K4047036</partinfo>, <partinfo>BBa_K4047037</partinfo>, <partinfo>BBa_K4047038</partinfo>). Its multiple components include an origin of replication (<partinfo>BBa_K4047029</partinfo>), a trc promoter (<partinfo>BBa_K4047032</partinfo>), an rrnB T1 terminator (<partinfo>BBa_K4047025</partinfo>), an rrnB T2 terminator (<partinfo>BBa_K4047026</partinfo>), a base of mobility region (<partinfo>BBa_K4047028</partinfo>), a lac repressor (<partinfo>BBa_K4047031</partinfo>), a lacIq promoter (<partinfo>BBa_K4047030</partinfo>), a lac operator (<partinfo>BBa_K4047033</partinfo>), and spectinomycin/streptomycin resistance (<partinfo>BBa_K4047027</partinfo>).  

Latest revision as of 18:59, 20 October 2021


pAM2991

The backbone sequence used for restriction digest assembly of our pGEM-FBPase, pGEM-tal, and pGEM-Tal/FBPase plasmids (BBa_K4047036, BBa_K4047037, BBa_K4047038). Its multiple components include an origin of replication (BBa_K4047029), a trc promoter (BBa_K4047032), an rrnB T1 terminator (BBa_K4047025), an rrnB T2 terminator (BBa_K4047026), a base of mobility region (BBa_K4047028), a lac repressor (BBa_K4047031), a lacIq promoter (BBa_K4047030), a lac operator (BBa_K4047033), and spectinomycin/streptomycin resistance (BBa_K4047027).

The backbone contains sequences homologous to the neutral site I region within Synechococcus elongatus PCC 7942, allowing target gene insertion without the disruption of other genes.

This backbone was able to successfully assemble with our target genes and create transformed cyanobacterial colonies, detailed on the MiamiU_OH 2021 wiki found at https://2021.igem.org/Team:MiamiU_OH

Although this backbone contains sequences that make it BioBrick incompatible, it is highly efficient for transformation in this cyanobacteria strain and contains registered components that may be useful in the assembly of other composite parts.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 7968
    Illegal XbaI site found at 2
    Illegal XbaI site found at 2546
    Illegal XbaI site found at 2860
    Illegal PstI site found at 14
    Illegal PstI site found at 2558
    Illegal PstI site found at 5013
    Illegal PstI site found at 5815
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 7968
    Illegal NheI site found at 2673
    Illegal PstI site found at 14
    Illegal PstI site found at 2558
    Illegal PstI site found at 5013
    Illegal PstI site found at 5815
    Illegal NotI site found at 515
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 7968
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 7968
    Illegal XbaI site found at 2
    Illegal XbaI site found at 2546
    Illegal XbaI site found at 2860
    Illegal PstI site found at 14
    Illegal PstI site found at 2558
    Illegal PstI site found at 5013
    Illegal PstI site found at 5815
  • 25
    INCOMPATIBLE WITH RFC[25]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 7968
    Illegal XbaI site found at 2
    Illegal XbaI site found at 2546
    Illegal XbaI site found at 2860
    Illegal PstI site found at 14
    Illegal PstI site found at 2558
    Illegal PstI site found at 5013
    Illegal PstI site found at 5815
    Illegal NgoMIV site found at 1043
    Illegal NgoMIV site found at 3454
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI site found at 198
    Illegal BsaI site found at 2068
    Illegal BsaI.rc site found at 4839
    Illegal BsaI.rc site found at 5605
    Illegal SapI site found at 3756